tbuckley/cabot-search huggingface dataset:
Abstract P1027: Cardioprotective Effects Of Genetically Engineered Cardiac Stem Cells By Spheroid Formation On Ischemic Cardiomyocytes
Authors: Han Saem Jeong, Jong-ho Kim, Soon Jun Hong
Journal: Circulation Research (2022)

Background: Sca-1+ cardiac stem cells and their limited proliferative potential were major limiting factors for use in various studies. Methods: Therefore, the effects of sphere genetically engineered cardiac stem cells (S-GECS) inserted with telomerase reverse transcriptase (TERT) were investigated to examine cardiomyocyte survival under hypoxic conditions. GECS was obtained from hTERT-immortalized Sca-1+ cardiac stem cell (CSC) lines, and S-GECS were generated using poly-HEMA. Results: The optimal conditions for S-GECS was determined to be 1,052 GECS cells/mm 2 and a 48 hours culture period. Compared to adherent-GECS (A-GECS) and S-GECS showed significantly higher mRNA expression of the growth factors SDF-1α and CXCR4. S-GECS conditioned medium (CM) significantly reduced the proportion of early and late apoptotic cardiomyoblasts during CoCl 2 -induced hypoxic injury; however, gene silencing via CXCR4 siRNA deteriorated the protective effects of S-GECS against hypoxic injury. As downstream pathways of SDF-1α/CXCR4, the Erk and Akt signaling pathways were stimulated in the presence of S-GECS CM. S-GECS transplantation into a rat acute myocardial infarction model improved cardiac function and reduced the fibrotic area. These cardioprotective effects were confirmed to be related with the SDF-1α/CXCR4 pathway. Conclusions: Our findings suggest that paracrine factors secreted from transplanted cells may protect host cardiomyoblasts in the infarcted myocardium, contributing to beneficial left ventricle (LV) remodeling after acute myocardial infarction (AMI).
Abstract P3084: T Helper Cell Polarity Determines Salt-sensitivity And Development Of Hypertension
Authors: InKyeom KIM, Jee Young Kim, Soyung Lee
Journal: Circulation Research (2022)

Background: Sodium chloride drives the stable induction of pathogenic T helper (Th) 17 cells and development of hypertension. However, high-salt intake induces hypertension in salt-sensitive persons but not in salt-resistant persons. Pathogenic Th17 cell plays an important role in the development of hypertension whereas suppressive Treg cell plays an important role in the prevention of hypertension. Objective: We tested the hypothesis that . Methods and Results: Eight-week-old male SS and SR rats were fed chow (NS, 0.4% NaCl) or high-salt (HS, 4% NaCl) diet for 4 weeks. Systolic blood pressure was measured by the tail-cuff method. Cultured splenic T cells from Dahl salt-sensitive (SS) and salt-resistant (SR) rats were exposed to hypertonic salt solution with or without IL-6 or TGFβ. T helper cell (Th17 and Treg) polarity was determined via flow cytometry. The expression of Th17-related genes such as interleukin (IL)-17A, IL-23R and retinoic acid receptor-related orphan receptor (ROR) γt) as well as Treg-related genes such as IL-10, CD25 and forkhead box (Fox) P3 were measured by ELISA or qRT-PCR. Adoptive transfer of Th17 cells isolated from SS or SR donors fed HS were performed intraperitoneally into SS and SR recipients. Upon chow diet, SR has higher Treg cells than SS does. HS intake induced hypertension and increased proinflammatory Th17 cells in SS but not in SR. Hypertonic salt solution induced differentiation of cultured splenic T cells preferentially into proinflammatory Th17 cells and anti-inflammatory Treg cells in SS and SR, respectively. Polarizing cytokine IL-6 also induced differentiation of cultured splenic T cells preferentially into proinflammatory Th17 cells and anti-inflammatory Treg cells in SS and SR, respectively, which were further potentiated by hypertonic salt solution. Adoptive transfer of activated Th17 cells isolated from hypertensive SS or normotensive SR, both of which had been fed HS diet, induced hypertension in SS recipients, but not SR recipients. Conclusions: These results indicate that T helper cell polarity after HS intake determines salt-sensitivity and development of hypertension in SS and SR rats. Th17 cell polarity predicts salt-sensitivity and development of hypertension, whereas Treg cell polarity predicts salt-resistance and maintenance of normotension. SR rats protect against hypertension after HS intake through anti-inflammatory Treg cell polarity.
Abstract P3002: Endothelial <i>Trpm2</i> Deletion Attenuates Ischemic Stroke By Preventing Blood-brain Barrier Disruption
Authors: Pengyu Zong, jianlin feng, Zhichao Yue, Zhichao Yue
Journal: Circulation Research (2022)

Introduction: Endothelial hyperpermeability is a hallmark of brain damage at the early stage of ischemic stroke. The subsequent blood-brain barrier (BBB) disruption drives the initial pathological changes and aggregates neuronal death. Ca 2+ overload is an important cause of endothelial dysfunction. TRPM2 is a Ca 2+ -permeable ion channel activated by oxidative stress, and oxidative stress is a critical contributor for endothelial hyperpermeability during ischemic stroke. Hypothesis: We hypothesized that TRPM2-mediated Ca 2+ influx plays a key role in causing endothelial hyperpermeability during ischemic stroke. Methods: Specific deletion of Trpm2 in endothelial cells was achieved by crossing Cdh5-cre mice with Trpm2 fl/fl mice. Middle cerebral artery occlusion (MCAO) was performed to simulate ischemic stroke. Tetrazolium chloride and Evans blue assays were used to evaluate infarct size and BBB leakage. Occludin and F4-80 / myeloperoxidase staining were used to detect tight junction degradation and immune cell invasion. Cerebral endothelial cells (CECs) were isolated and cultured. Oxygen-glucose deprivation (OGD) was used to simulate in vitro ischemic conditions. In vitro endothelial leakage assays were used to examine CEC permeability. Rhodamine-123 imaging and Ca 2+ imaging were used to evaluate oxidative stress and Ca 2+ overload in CECs. WB was used to examine CD36 signaling activation and whole-cell current recording was used to detect TRPM2 activation. Results: Specific deletion of Trpm2 in endothelial cells protects mice against MCAO-induced brain injury, which is characterized by reduced infarct size, mitigated plasma extravasation, reduced neutrophil / macrophage invasion and inhibited oxidative stress in the brain. In vitro experiments demonstrated that Trpm2 deletion alleviates ROS production, Ca 2+ overload and endothelial hyperpermeability induced by OGD and CD36 ligand thrombospondin-1 (TSP1). In CECs, activation of CD36 signaling induced by OGD and TSP1 was impaired by TRPM2 knockout. In transfected HEK293T cells, OGD and TSP1 induce the activation of TRPM2 currents in a CD36-dependent manner. Conclusion: Endothelial specific TRPM2 knockout attenuates ischemic stroke by preserving BBB integrity.
Abstract P1041: Redox Regulator MnTnBuOE-PyP5+ Protects Cardiomyocyte Against Oxidative Stress Mediated Myocardial Ischemia/reperfusion Injury
Authors: Sudha Sharma, Susmita Bhattarai, Hosne Ara, Utsab Subedi, Alika Shum, Foram Patel, Ines Batinic-Haberle, Hong Sun, Md Shenuarin Bhuiyan, Manikandan Panchatcharam, Sumitra Miriyala
Journal: Circulation Research (2022)

Background: Much evidence suggests that oxidative stress plays a major role in the pathogenesis of myocardial ischemia/reperfusion (I/R) injury and is a potential target for therapeutic interventions. We tested the hypothesis that the novel redox regulator, MnTnBUOE-2-PyP 5+ (BMX-001), mediates cardioprotection through reduction of oxidative stress in myocardial tissue following I/R injury. Methods and Results: BMX-001 (2mg/Kg, IP) or vehicle was administered in mice 4 hours before myocardial ischemia. Mice were then subjected to I/R injury by ligating the left anterior descending artery for 1 hour followed by 24 hours of reperfusion. Following 24 hours of reperfusion, infarct size, cardiomyocyte apoptosis, and SOD2 level were determined, and echocardiography was performed. Our study revealed that the redox regulator provided protection against I/R-induced injury in mice as evident by a significant reduction in infarct size, plasma LDH level, and oxidative stress. In addition, BMX-001 administration in mice markedly increased FoxO3a and SOD2 levels. BMX-001 treatment attenuated I/R induced cardiomyocyte apoptosis in-vivo and in-vitro as evident by a reduction in the number of TUNEL positive cells and improved cell viability. Echocardiography at 24 hours of reperfusion revealed improved ejection fraction and fractional shortening in mice treated with BMX-001. Furthermore, in-vitro treatment of BMX-001 (10μM) 24 hours before hypoxia/reoxygenation in H9c2 cells improved oxidative stress as evident by a reduction in cardiolipin peroxidation, mitochondrial superoxide level, and 4-HNE adducted proteins. Conclusion: Pre-treatment with redox regulator BMX-001 showed a beneficial effect in attenuating cardiomyocyte death, mitochondrial superoxide, and cardiolipin peroxidation following I/R injury in H9c2 cells. Pre-treatment with SOD2 mimetic improved heart function, decreased infarct size, and cardiomyocyte apoptosis in mice following I/R injury. Therefore, BMX-001 can be a novel cardioprotective agent against oxidative stress-induced myocardial I/R injury.
Abstract EC103: A Novel Nuclear Role For REDD1 In Cardiac Insulin Sensitivity And Resistance
Authors: Mason Wheeler, jessica Ibetti, Walter J Koch, Jessica M Pfleger
Journal: Circulation Research (2022)

Insulin resistance (IR) is a hallmark of type II diabetes (TIID) and causes cardiac dysfunction independent of hypertension and coronary artery disease, or diabetic cardiomyopathy (DC). Impaired metabolism and energetics are a major cause of DC, however the molecular mechanisms underlying these defects remain ill-defined. Our unbiased RNA sequencing (RNA-Seq) studies identified ‘regulated in development and DNA damage (REDD)1’ as a potential critical regulator of cardiac insulin sensitivity, as a 1.5-fold increase in REDD1 expression was observed in the hearts of mice treated with acute insulin (1 hour (h)) versus vehicle (n=3). REDD1 is an insulin-sensitive, negative regulator of mTORC1 and global REDD1 deletion leads to whole body IR. Interestingly, cardiac REDD1 is also upregulated in several models of IR. Thus, we hypothesized that while REDD1 is critical for cardiac insulin sensitivity, sustained elevated levels contribute to IR via chronic inhibition of mTORC1. Our findings confirm that acute insulin stimulation or chronic high fat diet (HFD) (2 to 16 weeks) induce cardiac REDD1 expression in vivo (1.7-fold±0.1, n=6). In neonatal rat ventricular myocytes (NRVM) or Hap1 cells, insulin (1 h) or palmitate (24 h) also induce REDD1 expression (1.3-fold±0.1, n=3 or 1.6-fold±0.1, n=9, respectively), which indeed inhibits mTORC1. Notably, insulin treatment following HFD or palmitate, does not further enhance REDD1. In addition, subcellular fractionation shows that REDD1 is uniquely detected in nuclear and chromatin fractions and increases with insulin (1.5-fold±0.1, n=6). We confirmed REDD1 nuclear localization via microscopy and chromatin binding via chromatin immunoprecipitation-deep sequencing (ChIP-Seq) (n=5, pooled). In addition, we determined that REDD1 primarily binds the transcription start sites of metabolic genes and increases with insulin, corresponding to reduced gene expression. Overall, our findings suggest that insulin-inducible cardiac REDD1 exists as part of negative feedback loops that prevent the overactivation of mTORC1 and metabolic gene expression, contributing to insulin sensitivity. Further, sustained fat-induced REDD1 expression, may drive IR via chronic inhibition of mTORC1 and metabolic gene expression.
Abstract P3071: Ikkε Deficiency Reduces P38 Activity Of Macrophages To Increases Cardiac Injury
Authors: Yong-sook Kim, Hyang H Cho, Dong-im Cho, Meeyoung Cho, Young Keun Ahn
Journal: Circulation Research (2022)

Background: Inhibitor of NF-κB kinase ε (IKKε) is a non-classical modulator of nuclear factor-kappa B (NF-κB), and is reported to regulate inflammation in various diseases. In this study, the involvement of IKKε in pathophysiological responses was examined in a mouse myocardial infarction model. Methods: MI was induced in IKKε-/- mice by coronary artery ligation. Neutrophils and macrophages were isolated from the heart tissues or bone marrows for further comparison studies by Western blot, PCR, and flow cytometry. Cell phenotypes were identified by immunofluorescence staining. Results: The IKKε-/- group showed poor early survival rate, high circulating IL-6 level, large cardiac fibrosis, and low ejection fraction (30.33±5.25% vs. 34.02±5.53%, p&lt;0.05) compared with the wild type group. Further study revealed that the numbers of neutrophils and macrophages in the heart were higher in the IKKε-/- group than in the PBS group. Macrophage phenotype was more inflammatory along with lower p38 activity in the IKKε-/- group. Importantly, we identified macrophage-myofibroblast transition (MMT) in the infarcted myocardium and found that the number of MMT F4/80(+)SMA(+) cells was 2-fold higher in the IKKε-/- group. The inactivation of p38 accelerated MMT in human macrophages, and the level of phosphorylated p38 was lower in infarct zone of human failing heart tissues. Conclusion: Collectively, IKKε-p38 axis may lead macrophages to aggravated inflammation and accelerated MMT, resulting in poor outcome after myocardial infarction. Here, we suggest that restoring IKKε or p38 in macrophages could be the target to attenuate cardiac inflammation.
Abstract P3018: A Requirement For P21 Activated Kinase1 To Preserve Cardiac Function And Metabolic Homeostasis
Authors: Paola C Rosas, Ashley Batra, Koreena Rafael-Clyke, Shreya Kurup, Shamim Chowdhury
Journal: Circulation Research (2022)

Background/Hypothesis: Heart failure with preserved ejection fraction (HFpEF, EF&gt;50%) continues to lack effective treatment. Obesity and diabetes are strong predictors of HF, with &gt;80% of HFpEF patients being obese or overweight and ~40% suffering from diabetes. P21 activated kinase-1 (PAK1) is a member of the highly conserved family of serine/threonine protein kinases regulated by Cdc42/Rac1. We tested the hypothesis that PAK1 is an important element in the regulation of cardiac function and metabolic homeostasis. Material and methods: 3-6 months male and female global PAK1 knock-out mice (PAK1 -/- ) and wild type (WT) controls were used to measure % of fat mass (by NMR), adipocyte size and glucose tolerance. We used echocardiography to assess cardiac dysfunction and western blots (WB) to examine cardiac protein alterations. Results (see Table): PAK1 -/- hearts exhibited preserved ejection fraction and diastolic dysfunction with slower e’, increased E/e’ ratio and prolonged IVRT. Additionally, PAK1 -/- hearts showed dilated left atrium and thickening of posterior wall. WB analyses showed decreased phosphorylation of cMyBP-C and dysregulation of calcium handling proteins. Moreover, PAK1 -/- mice exhibited increased % fat mass, adipocyte hypertrophy and impaired glucose tolerance test. Conclusion: Deletion of PAK1 resulted in obesity, impaired glucose homeostasis and HFpEF. Thus, PAK1 is required to support cardiometabolic health.
Abstract P3013: MicroRNA-122 Targeting With Gamma Peptide Nucleic Acid Inhibitor Improves Vascular Endothelial Dysfunction In Pre-diabetic Mice.
Authors: Ravinder R Gaddam, Karishma Dhuri, Yung-Rae Kim, Julia S Jacobs, Vikas Kumar, Qiuxia Li, Kaikobad Irani, Raman Bahal, Ajit Vikram
Journal: Circulation Research (2022)

Background: In patients with diabetes, the endothelial function precedes and predicts the risk of cardiovascular disorders. Current treatment approaches fail to decrease unwanted cardiovascular outcomes in these patients. The blood levels of microRNA-122 (miR-122) elevated in patients with diabetes and correlated with risk and severity of cardiovascular disorders. Therefore, targeting miR-122 with efficient and safer inhibitors could be a promising approach. The objective of this study was to investigate the effectiveness of gamma peptide nucleic acid (γPNA)-based miR-122 inhibitor in preventing the development of endothelial dysfunction in pre-diabetic mice. Methods: To test the effectiveness of γPNA technology in inhibiting miR-122, we synthesized γPNA-miR-122 inhibitor (γP-122-I) and γPNA-scrambled control (γP-SC). The purity and molecular weight of γP-122-I and γP-SC were assessed by HPLC and MALDI spectrometry, respectively. The binding and affinity of γP-122-I were determined by gel-shift assay and thermal denaturation. The insulin-resistance (induced by high-fat-diet, HFD) mice were injected with γP-122-I (5 mg/kg/day, i.p., 6 weeks). miR-122 expression was measured by real-time PCR. The safety of γP-122-I was evaluated by biochemical and histological analysis of vital organs (liver &amp; kidney). The vascular endothelial function was assessed by determining endothelium-dependent and independent relaxation of phenylephrine-induced precontracted aortic rings. Results were expressed as mean ± s.e.m. Results: The HPLC of γPNA oligomers show that both γP-122-I and γP-SC were &gt;95% pure. The molecular weight of γP-122-I (m/z: 9227.4) and γP-SC (m/z: 9232.7) were confirmed by MALDI spectrometry. γP-122-I strongly binds with miR-122 and forms a stable complex compared to commercially available miR-122 inhibitor. A comprehensive safety analysis of γP-122-I in mice shows that neither it affected liver and kidney function nor their morphology during acute and chronic exposure, respectively. Next, HFD-fed mice show higher serum and aortic miR-122 levels, endothelial dysfunction, and inflammation, which were reversed by γP-122-I treatment. Conclusions: γP-122-I inhibits the development of impaired endothelial function in pre-diabetic mice without any evidence of toxicity
Interexpert Agreement Between Emergency Neuroradiologists with Different Levels of Experience in the Rating of ASPECTS
Authors: P. L. Andropova, P. V. Gavrilov, I. P. Kazantseva, N. I. Kochanova, A. N. Narkevich, T. N. Trofimova
Journal: Radiology - Practice (2022)

Fifteen radiologists independently evaluated fifty non-contrast brain computed tomography of patients with middle cerebral artery (MCA) stroke. Within-group agreement was either negligible or moderate, and the best inter-expert agreement was observed between neuroradiologists with 1 to 3 years of experience. Our study allows us to conclude that ASPECTS is not an ideal tool for standardizing the diagnosis of MCA stroke, therefore, it is necessary to develop new models of standardization systems, as well as the use of hydride diagnostics, including automatic analysis.
Abstract P3047: Identification Of Role Of Intracellular Chloride Channel Clic4 In Diabetic Cardiomyopathy.
Authors: Devasena Ponnalagu, Katherine Wright, Diego Antelo, Sahej Bindra, Kristin I Stanford, John C Edwards, Harpreet Singh
Journal: Circulation Research (2022)

Cardiovascular diseases are the most common cause of morbidity and mortality in Diabetic patients. Mitochondrial dysfunction is the major contributor of the myocardial structural, functional, and metabolic defects in diabetic cardiomyopathy. Activation of mitochondrial ion channel, K ATP have been shown to facilitate protection from the adverse effects of diabetic cardiomyopathy via modulating mitochondrial function. In this study we delineate the impact of mitochondrial associated membrane (MAM) chloride channel-CLIC4 in diabetic cardiomyopathy. Our results show that pharmacological inhibition of chloride intracellular channel (CLIC) with IAA-94 abrogated the cardio-deleterious effects of hyperglycemia in neonatal cardiomyocytes, thus, implicating role of targets of IAA-94 in the onset and development of diabetic cardiomyopathy. Amongst the six mammalian paralogs of CLICs, we identified CLIC4 and CLIC5 are most abundant in the heart. In addition, we observed over expression and localization changes of MAM-CLIC4 but not mitochondrial CLIC5 in neonatal cardiomyocytes upon high glucose treatment. Furthermore, cardiomyocytes isolated from CLIC4 knock out mice under hyperglycemia do no exhibit disruption of sarcomere structures, reduction in contractility, and enhanced ROS production in comparison to wild type cardiomyocytes. These observations suggest that an absence of CLIC4 revokes the structure-function changes in cardiomyocytes upon hyperglycemia. Moreover, we demonstrate that clic4 -/- mice were protected from body weight gain and impaired glucose tolerance in comparison to wild type mice in the presence of a high fat diet. Interestingly, we also observed that clic4 -/- mice are resistant to increase in blood glucose levels upon streptozocin administration. Thus, suggesting a regulatory role of CLIC4 in glucose metabolism. Overall, this study suggests a probable novel role of intracellular chloride channel CLIC4 in diabetic cardiomyopathy via regulating glucose metabolism.
Abstract P3094: Mir-181c-5p As Detrimental Player In A Mouse Model Of Cardiorenal Syndrome With Thrombotic Microangiopathy
Authors: Jente R Boen, Dustin Krüger, Amélie Dendooven, Tine Bruyns, Jens Van Fraeyenhove, Eline Feyen, Andreas B Gevaert, Vincent F Segers, Emeline M Van Craenenbroeck
Journal: Circulation Research (2022)

Background: MiR-181c-5p is described to induce heart failure (HF), while its role in renal pathology is undetermined. Renal dysfunction is present in 40-60% of HF patients and associated with poor prognosis. This study is the first of its kind to investigate the role of miR-181c-5p in a mouse model of cardiorenal disease (CRD). Our hypothesis states a protective effect of miR-181c-5p inhibition on HF development. Methods: CRD was induced by feeding male C57BL/6J mice (n=20) a high-fat diet (HFD) and L-NAME in drinking water (5g/L) for 6 weeks, angiotensin-II was co-administered via osmotic minipumps (1000ng/kg/min) during the final 2 weeks. Healthy controls (n=16) underwent sham-surgery. Mice were randomly assigned to weekly injections (40mg/kg) with miR-181c-5p antagomiR (INH) or scrambled control for the duration of the study. We assessed cardiac function (echocardiography, hemodynamics), renal function (plasma creatinine), target expression (RT-qPCR), and histology. Results: CRD animals showed mild systolic and diastolic cardiac dysfunction compared to healthy controls, with cardiac fibrosis (2.5±0.3 vs 1.8±0.2% area; p=0.0004) and hypertrophy (0.11±0.03 vs 0.08±0.01g/cm; p=0.005). Renal dysfunction presents with kidney atrophy (0.07±0.006 vs 0.09±0.01g/cm; p=0.02), increased plasma creatinine (21±6 vs 10±5; p=0.01), renal fibrosis (0.26±0.22 vs 0.005±0.21 % area; p=0.036) and glomerular abnormalities (glomerulosclerosis, hypertrophy/atrophy, mesangial matrix expansion, reduced podocyte number). CRD+INH animals had comparable cardiac phenotype to CRD (p&gt;0.05). Their renal phenotype was exacerbated with elevated glomerular damage (26±3 vs 18±9;p=0.04) and significantly increased mortality rate (50%) (Kaplan-Meier p=0.01) compared to healthy controls (0%) or CRD (20%), associated with increased occurrence of tubular atrophy, endothelial swelling and systemic thrombotic microangiopathy (TMA). qPCR analysis identified Vegf as potential target of miR-181c-5p. Conclusion: This study demonstrates a detrimental effect of miR-181c-5p inhibition on renal function in a CRD mouse model, driven by glomerular damage and TMA through Vegf signaling. Cardiac function was unaffected.